Effects of N-Arachidonoylethanolamine on the Quality of Storage Platelets

Abstract

Objective To evaluate the impact of N-Arachidonoylethanolamine(ANA)on the quality of platelets(PLT)stored at(22±2)℃.Methods From November 7th to 25th 2011,a total of 20 volunteers who donated PLT in Blood Center of Shandong Province were included into this study.Apheresis PLTs were collected through the Amicus instrument.Different working concentrations of ANA(0.1,0.5,1,5,10 and 50 μmol/L)were added to the PLTs samples,respectively,as the experimental group,another corresponding parts with no ANA as the controls,respectively.All the PLT samples were stored at(22±2)℃ for 7 days,then were evaluated the viability by measuring the mitochondrial reduction with methyl thiazolyl tetrazolium colorimetric for detecting the most effective concentration of ANA.Samples(50 mL/each)were taken and split into two storage parts(25 mL/each).The most effective concentration of ANA was added one part as the experimental group(n=20),the other with no ANA as the control group(n=20).These PLT samples were stored on a flat-bed shaker at(22±2)℃.PLT count,mean PLT volume (MPV),PLT distribution width(PDW),phosphatidyl serine(PS)and soluble P-selectin of the two groups were detected on the 1st,3rd,5th,7th,9th and 11th day of storage.Results The most effective final concentration of ANA was 0.5 μmol/L,which showed significant increasing PLTs survival(about 1.7 fold more than controls)(t=14.32,P＜0.05).During 11 days of storage,the PLT counts were not changed significantly until on the 11th day of storage between the experimental group and control group.Moreover,the MPV and PDW were decreased significantly in the experimental group than those in the control group on the 11th day of storage(P＜0.05),respectively.The rates of PLT PS positive were on the rise during the storage period:the experimental group were from(8.29 ±1.44)％ to(21.18 ± 1.78)％ while the control group were from(14.24±2.47)％ to(40.6± 1.95)％,with significant differences between the two groups (P＜0.05)on the 7th,9th and 11th day of storage,respectively.Soluble P-selectin contents of the experimental group on the 7th,9th and 11th day of storage were(75.08± 6.35)ng/mL,(81.58±6.09)ng/mL and(103.55±7.91)ng/mL,while those in the control group were(90.37±8.91)ng/mL,(117.56 ± 9.98)ng/mL and(144.7 ± 7.76)ng/mL,respectively,with significant differences between the two groups(P＜0.05).Conclusions 0.5 μmol/L ANA had a good protective effect on PLTs,and could potentially alleviate PLT storage lesions to some extent. Key words: endocannabinoids; blood platelet; blood preservation